拟南芥应答非生物胁迫的直接质谱分析研究

Metabolomics mainly investigates the change of endogenous metabolism of biological system that is stimulated or disturbed, and studies the metabolic pathways of biological system. Following genomics, transcriptomics and proteomics are other important fields of researches of systems biology. Metabolomics focuse on the variations of small molecular (MW＜1000) of metabolism circulation, and reflect changes in the metabolomics for response of the cells or tissues, which are disrupted by outside stimuli or genetic modification.The study of metabonomics for Arabidopsis thaliana response to abiotic stress is the key to master tolerance and sensitivity of plant response to abiotic stress, which is complex in life process. Our self-developed neutral desorption extractive electrospray ionization mass spectrometry (ND-EESI-MS) is a new type of direct ionization technology. Several advantages of EESI-MS as follows: 1) ND-EESI can be used for direct analysis of samples without preparation; 2) ND-EESI tolerates complex matrices and it has good stability and high sensitivity; 3) ND-EESI has a wide range of applications and can be used to detect the polar and non-polar substances via ion/molecule reactions; 4) the sample object is not exposed to electric field or to chemical contamination, so ND-EESI has potential applications for biological samples, such as, chemical reaction monitoring, real time and remote analysis of plants or animals; 5) ND-EESI can be coupled with a variety of instruments, and it is easy for miniaturization and integration; 6) selective molecule-ion or ion-ion reactions can be easily implemented in ND-EESI. This project adopts ND-EESI-MS to analyse mass spectrometric of metabonomics for Arabidopsis thaliana response to abiotic stress directly, and to establish a qualitative, quantitative, in situ and dynamic analysis method of Arabidopsis thaliana metabolite. Through comparing fingerprint spectra of metabolomics under different abiotic stress conditions, meanwhile, extracting and selecting iconic compounds in metabolites related to the abiotic stress conditions, to investigate the relationship between metabolomics of Arabidopsis thaliana response to abiotic stress and the corresponding conditions. Consequently, we establish relevant mathematical modele, and metabolic pathway of Arabidopsis which responses to the typical abiotic stress. Based on the direct mass spectrometry analysis, we hope to develop a new research method of plant metabolomiacs, and to provide a new technology and idea for Botanical research.

植物应答非生物胁迫代谢组学研究，是理解植物对非生物胁迫耐受性和敏感性复杂生命过程的关键。中性解吸电喷雾萃取电离质谱（ND-EESI-MS）技术是新兴的直接离子化技术。它无需样品预处理即可对复杂基质生物样品进行快速质谱分析，且在电离过程中，待测样品不会与高电压直接接触、不受化学污染和物理损伤，是一种可对活体样品进行原位分析，且不会附带产生额外胁迫作用的快速检测新技术。本项目拟采用ND-EESI-MS技术，对拟南芥应答非生物胁迫的代谢产物进行直接质谱分析，建立其代谢产物定性/定量分析方法；通过对比不同胁迫条件下拟南芥代谢组学的指纹谱图，结合传统的分析检测方法，提取并筛选出与胁迫条件相关的标志性代谢产物，探讨胁迫条件与拟南芥应答代谢之间的关系，并基于此建立相关数学模型；构建拟南芥对典型非生物胁迫环境应激反应的代谢途径。开发基于直接质谱分析的植物代谢组学研究方法，为植物学研究提供新技术和新方法。

干旱是温室效应导致的全球气候变化重要的表征，严重影响了农作物的分布、产量。植物应答非生物胁迫代谢组学研究，是理解植物对非生物胁迫耐不同层次调控网络的关键。如何高效、快速从植物组织这样复杂基质中获取整体样品中与胁迫相关的代谢组成分成是本研究的前提。本项目拟采用ND-EESI-MS技术，对拟南芥应答非生物胁迫的代谢产物进行直接质谱分析，建立其代谢产物定性/定量分析方法；通过对比不同胁迫条件下拟南芥代谢组学的指纹谱图，结合传统的分析检测方法，提取并筛选出与胁迫条件相关的标志性代谢产物，探讨胁迫条件与拟南芥应答代谢之间的关系，并基于此建立相关数学模型；构建拟南芥对典型非生物胁迫环境应激反应的代谢途径。开发基于直接质谱分析的植物代谢组学研究方法，为植物学研究提供新技术和新方法。具体内容如下：.1）构建了基于EESI-MS、DAPCI-MS、iEESI-MS的拟南芥直接质谱分析平台；.2）采用EESI-MS对高温高湿胁迫下咖啡种子活力进行了评价；.3）采用EESI-MS对高温高湿胁迫下玉米种子活力进行了评价；.4）采用DAPCI-MS对高温高湿胁迫下咖啡种子活力进行了评价；.5）采用DAPCI-MS对高温高湿胁迫下玉米种子活力进行了评价；.6）采用iEESI-MS进行了PEG6000胁迫小白菜代谢产物的分析；.7）采用iEESI-MS进行了铀胁迫水葫芦代谢产物的分析；.8）采用 EESI-MS研究了水葫芦对铀的去除作用。.在本项目的支持下，发表论文6篇，其中SCI收录论文5篇，达到项目考核指标。